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Site-specific bacterial chromosome engineering mediated by IntA integrase from Rhizobium etli

Overview of attention for article published in BMC Microbiology, June 2016
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Title
Site-specific bacterial chromosome engineering mediated by IntA integrase from Rhizobium etli
Published in
BMC Microbiology, June 2016
DOI 10.1186/s12866-016-0755-y
Pubmed ID
Authors

Rogelio Hernández-Tamayo, Gonzalo Torres-Tejerizo, Susana Brom, David Romero

Abstract

The bacterial chromosome may be used to stably maintain foreign DNA in the mega-base range. Integration into the chromosome circumvents issues such as plasmid replication, stability, incompatibility, and copy number variance. The site-specific integrase IntA from Rhizobium etli CFN42 catalyzes a direct recombination between two specific DNA sites: attA and attD (23 bp). This recombination is stable. The aim of this work was to develop a R. etli derivative that may be used as recipient for the integration of foreign DNA in the chromosome, adapting the IntA catalyzed site-specific recombination system. To fulfill our aim, we designed a Rhizobium etli CFN42 derivative, containing a "landing pad" (LP) integrated into the chromosome. The LP sector consists of a green fluorescent protein gene under the control of the lacZ promoter and a spectinomycin resistance gene. Between the lacZ promoter and the GFP gene we inserted an IntA attachment site, which does not affect transcription from the lac promoter. Also, a mobilizable donor vector was generated, containing an attA site and a kanamycin resistance gene; to facilitate insertion of foreign DNA, this vector also contains a multicloning site. There are no promoters flanking the multicloning site. A biparental mating protocol was used to transfer the donor vector into the landing pad strain; insertion of the donor vector into the landing pad sector via IntA-mediated attA X attA recombination thereby interrupted the expression of the green fluorescent protein, generating site-specific cointegrants. Cointegrants were easily recognized by screening for antibiotic sensitivity and lack of GFP expression, and were obtained with an efficiency of 6.18 %. Integration of foreign DNA in Rhizobium, lacking any similarity with the genome, can be easily achieved by IntA-mediated recombination. This protocol contains the mating and selection procedures for creating and isolating integrants.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 17 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United Kingdom 1 6%
Unknown 16 94%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 5 29%
Researcher 4 24%
Student > Master 2 12%
Student > Doctoral Student 2 12%
Lecturer > Senior Lecturer 1 6%
Other 2 12%
Unknown 1 6%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 6 35%
Agricultural and Biological Sciences 5 29%
Veterinary Science and Veterinary Medicine 2 12%
Chemistry 1 6%
Unknown 3 18%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 01 July 2016.
All research outputs
#14,856,117
of 22,879,161 outputs
Outputs from BMC Microbiology
#1,602
of 3,195 outputs
Outputs of similar age
#212,790
of 352,012 outputs
Outputs of similar age from BMC Microbiology
#47
of 92 outputs
Altmetric has tracked 22,879,161 research outputs across all sources so far. This one is in the 33rd percentile – i.e., 33% of other outputs scored the same or lower than it.
So far Altmetric has tracked 3,195 research outputs from this source. They receive a mean Attention Score of 4.1. This one is in the 44th percentile – i.e., 44% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 352,012 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 36th percentile – i.e., 36% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 92 others from the same source and published within six weeks on either side of this one. This one is in the 43rd percentile – i.e., 43% of its contemporaries scored the same or lower than it.